Laboratory Animal Science

Murine Subcutaneous Transplantation Model of Hepatoma with Hepa1-6 Cell

ZHANG Zhi-deng~1,CAO Zhi-yun~2

2009, 26(04): 1-3.

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Objective To develop a murine subcutaneous transplantation model of hepatoma with cell line Hepa1-6.Methods Hepa1-6 cell was cultured in vitro and its growth was monitored;Hepa1-6 cells were injected intradermally into C57BL/6J mice at concentrations of 2×10~5/mice(A group),2×10~6/mice(B group)and 4×10~6/mice(C group).Results The cells grew very slowly,and they went into exponential growth three days after assimilation;the rate of tumorigenesis in group A,B and C was 30%,80% and 100% respectively;Tumorigenesis was observed after 12 and 10 days in group A and B respectively,but after 7 days in group C;From the third to the forth week,the tumor bulks in group C and B grew significantly bigger than those in group A(P<0.01). There was no obvious difference in tumor bulks between Group C and B.Conclusion The murine hepatoma model can be established by subcutaneous transplantation of Hepa1-6 cells at the concentration of 4×10~6/mice.

Effects of Ginseng Stem Leaf Glucoside on Mouse Arrhythmia and Death Time

TANG Ze-yao~1,TANG Tian-tian~1,FU Lei~2,TANG Yu-yi~1,XIANG Yuan-yuan~1,FAN Hong-bo~1,LIN Yuan~1

2009, 26(04): 4-7.

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Objective To investigate the effect of Ginseng stem leaf glucoside on arrhythmia in laboratory mice.Methods Mouse arrhythmia was induced by chloroform or aconitine,and the effect of Ginseng stem leaf glucoside on anti-arrhythmia was examined.Results Ventricular arrhythmia induced by chloroform or aconitine was inhibited and the death time was delayed by Ginseng stem leaf glucoside in a dose dependent manner.Conclusion Ginseng stem leaf glucoside can antagonize arrhythmia induced by chloroform or aconitine.

Quantitative Assay of Atherosclerotic Lesion in Mouse Aortic Root

QIAO Zheng-guo~1,YAN Zhong-qun~2,REN Jing-yi~1,CHEN Hong~1

2009, 26(04): 8-11.

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Objective To describe a novel method for quantifying the volume of mouse atherosclerotic lesion using the ApoE-knockout mice.Methods Frozen sections were cut from the aortic roots of the 16-year old female ApoE-knockout mice using the standardized cryosection protocol.Seven serial sections were collected at 100 μm intervals and stained with oil red-O.Lesion area and volume were analyzed and calculated using Leica Q500MC image analysis software and Cavalieri formula respectively.Results The average atherosclerotic area and lesion volume were(520 632) μm~2 and(364 442) μm~3 respectively.Conclusion The introduced method is easy to be operated and standardized.

Study of the Rat Model of Huntington's Disease Induced by 3-Nitropropionic Acid

LI Feng,XU Qun-yuan

2009, 26(04): 12-15.

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Objective To evaluate the efficiency of two different administrations of 3-Nitropropionic acid(3-NPA) for inducing rat model of Huntington's disease.Methods Twelve SD rats were divided into two groups,one receiving continuous administration of 3-NPA(15 mg/kg daily,ip,for 5 days) and the other receiving intermittent administration(15 mg/kg daily,ip,for 2 days,no treatment for 2 days,and then 15 mg/kg daily,ip,for 3 days).Results Mortality rate was decreased to 25% and the degenerating cell bodies and affected fibers in striatum were found rapidly by Fluro-Jade B staining in the intermittent administration group.Conclusion The intermittent administration of 3-NPA is an effective method for inducing animal model of Huntington's disease.

Experiences in Creating Four Types of Acute Renal Failure Models

LIU Xiang~1,ZHANG Bai-yu~2,WANG Rong~1

2009, 26(04): 16-19.

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To introduce the operation,pathogenic mechanism,pathological changes and clinic courses of four types of acute renal failure(ARF) models.Animal ARF can be induced by renal ischemia-reperfusion injury,glycerine,gentamicin and mercuric chloride,and typical acute tubular necrosis can be observed in pathological changes.Different ARF models show different etiopathogenisis,pathological changes and clinic courses,so they could be chosen depending on different requirements in practical work.

Effect of Nasal Aerosol on Behavior and Symptom of the Migraine Rat Model Induced by Nitroglycerin

ZHONG Rong-ling,HUANG Hou-cai,YANG De-gong,CAO Peng

2009, 26(04): 20-22.

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Objective To investigate the effect of nasal aerosol on the behavior and symptom of experimental migraine rat model.Methods Sixty SD rats were divided randomly and evenly into 6 groups: blank control group,model group,positive drug group and 3 treatment groups.The treatment groups included the model rats which were administrated with high,middle and low doses of nasal aerosol respectively;the positive drug group received gastric perfusion of Tiansu,and distilled water was given by gastric perfusion to the model group and the blank control group.Behavior and symptom changes of the rat models were observed continuously at various time intervals.Results The behavior and symptom of treatment groups were significantly better than that of model group.Conclusion The nasal aerosol improves the behavior and symptom of migraine rats greatly.

Effect of Hemoperfusion Adsorption in Dog Model of Endotoxemia

ZHAO Shi-hai~1,WANG Li~2,HUANG Xiao-qiong~1,ZHONG Zhi-yong~1,SUN Xia~1,WANG Gang~1

2009, 26(04): 23-27.

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Objective To explore the efficacy and safety of hemoperfusion adsorption in dogs.Methods Beagle dog model of endotoxemia was established by intravenous infusion of endotoxin LPS,and was randomly divided into treatment group and control group respectively.Treatment group was given the perfusion adsorption therapy using hemoperfusion machine and endotoxin adsorption column,and the control group was treated with the non-endotoxin adsorbent column.Blood physiological and biochemical parameters and endotoxin content were measured at multiple time points.Results Safety tests: body temperature dropped continuously in the treated dogs,but rose at first and then fell in the control dogs.In 90 min and 120 min after perfusion,body temperature was significantly different between the control and treatment group(P<0.05).Compared with the control group,there were no significant differences in heart rate,blood routine,serum total protein and albumin,electrolyte concentration, prothrombin time,activated partial prothrombin time,serum complement C3 concentration,thrombin content,and so forth(P>0.05).Detection of adsorption efficiency: the level of serum endotoxin was decreased significantly in the treatment group(P<0.01).Conclusion Serum endotoxin can be adsorbed and the level of serum endotoxin can be reduced by the endotoxin adsorption column,which is of good blood compatibility.

Quality Control of the Laboratory Zebrafish and Acute Toxicity Test on the Zebrafish in Different Health Conditions

ZHANG Jing-ji,YIN Hao-wen,ZHAO Hua-qing

2009, 26(04): 28-31.

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Objective To study the toxicity of zebrafish in different health conditions exposed to the same chemical pollutant,and to discuss the suitability of current quality control(QC) of laboratory fish. Methods According to the OECD 203 Fish Acute Toxicity Test method,healthy and sick zebrafish were placed in methomyl and potassium dichromate(K_2Cr_2O_7) respectively.Results and Conclusion The values of K_2Cr_2O_7 24 h-LC_(50) and Methomyl 96 h-LC_(50) of the healthy fish were 323.38 mg/L and 4.91 mg/L respectively,and were 302.55 mg/L and 3.04 mg/L in the sick fish.Although the K_2Cr_2O_7 24 h-LC_(50) values of both fish meet the QC requirement,there is a significant difference on the 96h-LC_(50) values between the two groups of fish in the acute toxicity test of methomyl.

Mini-Pig Model of Gastrohelcoma Induced by Different Irritant Reagents

TIAN Yu-ying,LUO Ming-fu,XU Yi-hui,ZHANG Wei-bo,WANG Guang-jun,WANG Rui-hong,LI Cui-hong

2009, 26(04): 32-34.

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Objective Establishing mini-pig model of gastrohelcoma.Methods Three different irritant reagents were injected into the stomach of mini-pigs through gastric tube and the tissue of stomach was gotten after killing the pig three hours after the injection.The tissue was embedded by paraffin,sliced and dyed by HE which was observed and photo under microscope.Results After injecting 95% alcohol,the gastric mucous membrane was intact,light red and smooth while only a little congestion,edema and deep red color in loci under macrography.The epidermis and gastric foveola was intact while swelling and slightly damaged in loci under microscope.After injecting anhydrous alcohol,although the gastric mucous membrane was still intact around pylorus,but the membrane with red color was deeper and wider than that of injecting 95% alcohol.Under microscope,the epidermis was damaged and became obviously thin but still some epidermis was kept.The gastric foveola became short and gastrohelcoma appeared obviously.After injecting 36% glacial acetic acid,the internal wall of stomach hunched with white and yellow color which was ischemia and dead in tissue.The epidermis was damaged severely and disappeared partly while inherent membrane tissue appeared.Conclusion injecting glacial acetic acid through gastric tube is ideal method to make gastrohelcoma.

Application of the E-Net System on Temperature and Humidity Monitoring in Conventional Animal Laboratory

JIN Shu-xing~1,SHI Guo-chun~2

2009, 26(04): 35-40.

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The conventional animal laboratory of Zhengzhou University was established in 2005,and the room temperature was controlled by multi-groups of small split central air-conditioners.To meet the GLP requirements,a remote temperature and humidity monitoring system based on multi-CPU was designed and applied without changes in the original system.The advanced single-chip technology was applied in temperature and humidity recorder,which is of high degree of intelligence and data precision.Independent control system in each region or room achieved temperature and humidity control or alarm,and real-time and remote control of other devices can be operated through the extending interface.This system has the capabilities of automatic recording,control,and data management,but installation and management of the system is easy.Using the conventional computer,remote communication and multi-point monitoring can be achieved.This system is stable and reliable so that investments will be saved significantly.It can be commonly used and promoted widely.

Laboratory Animal Science in Zhejiang Province

ZHU Xiao-feng

2009, 26(04): 41-44.

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Compared with the advanced international and domestic levels,laboratory animal science has not been considerably developed in Zhejiang province.The main task for laboratory animal science in Zhejiang province is to establish the public service platform consisted of production and supply,application research,quality control and personnel training systems.The key work in the future is also proposed in this review.

The Common Diseases and Main Problems for Research of Standardization of the Laboratory Zebrafish

WU Si-ao~1,WANG Qing~1,GAO Chang~2,MU Yuan~2,ZHAO De-ming~1,SUN Deming~2

2009, 26(04): 51-56.

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Zebrafish is a kind of fish widely used in laboratories.The standardization of laboratory zebrafish has not been established and there is even no national or regional approved technique standard.The purpose of this review is to collect and analyze the information of the common diseases of zebrafish and the possible problems that may occur in the procedure of biological purification,which will provide the technical basis for formulation of laboratory fish standardization and quality control,and for evaluation of standarlized zebrafish species.

Effect of Three Anesthetic Drugs in SD Rat Liver Transplantation Surgery

CHEN Hui,JIANG Li,WANG Yan,YU Da

2009, 26(04): 57-59.

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Objective To compare the anesthetic effect of ketamine hydrochloride,pentobarbital sodium,and trichloro acetaldehyde hydrate in SD rat liver transplantation surgery,and selected the suitable drugs and doses to rats liver transplantation.Methods Sixty SD rats were divided randomly into three groups treated with the three anesthetic drugs respectively.The anesthetic efficiency,respiratory excretion,body temperature,heart rate and respiratory rate were compared,and the low dose of anaesthetic method was found to be the best.Liver cirrhosis was induced in 102 SD rats,which were anaesthetized using the preferable anaesthetic method and used in liver transplantation surgery.Results All of the three anesthetic drugs were effective,but different in terms of the complexity of the operation,the duration of anesthetic effect,and the adverse reaction.Rats waked up quickly and recovered better when low dose of pentobarbital sodium or trichloro acetaldehyde hydrate was given in the liver transplantation surgery.Conclusion In a complex surgery,such as rat liver transplantation,it is necessary to make the animal wake up quickly using the preferable anesthetic drugs and dosages and the low dose of anaesthetic method could get the good results.

Detection of Antibodies Against B Virus in Malaysia Crab-Eating Macaques(Macaca fascicularis)

LIU Huan~1,ZHANG Wei~

2009, 26(04): 60-62.

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Objective To investigate the positive rate of antibodies against B virus in Malaysia Crab-eating Macaques(Macaca fascicularis).Methods Blood were collected from the femoral veins of total 1916 Macaca fascicularis captured in the wild,then sera were isolated and examined using the immune enzymatic slide(EIA) and ELISA diagnostic kit.Results The total positive rate of antibodies against the B virus was 68.5%.For the monkeys below 2 years,2 to 4 years and 4 to 6 years old,positive rate was 53.0%,63.8% and 76.4% respectively.Conclusion The positive rate of antibodies against B virus is high and increases with aging in Macaca fascicularis.Infection of B virus is not influenced by gender.

A Scientific Bedding-Changing Procedure by Replacing Wood-Shavings with Corn Cob

LI Bao-wen,LIANG Chun-nan,LI Shao-ming

2009, 26(04): 65-68.

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Objective To develop a scientific bedding-changing procedure to replace the wood shavings with corn cob.Methods Two groups of SD rats(weight about 500 g,n=72 and 200 g,n=24,respectively) and 1 group of(BALB/c) mice(weight 35 g,n=24) were used,and ammonia concentration,hygroscopicity of bedding as well as bedding and fur cleanliness under different conditions were evaluated.Results and Conclusion The present study indicates that volumetric absorbencies of corn cob con ammonia is better than that of wood shavings,and the cages bedded with corn cob keep a longer bedding-changing interval.

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